活性氧清除剂 NAC 抑制阴道毛滴虫排泄分泌物诱导的 SiHa 细胞凋亡

本研究旨在探讨活性氧（ ROS）清除剂N－乙酰－L－半胱氨酸（ NAC）能否抑制滴虫排泄分泌物（ Tv ESP）诱导的人子宫颈癌SiHa细胞凋亡。体外培养阴道毛滴虫并制备Tv ESP,分别用100μg／mL Tv ESP和PBS处理SiHa细胞, Tv ESP处理SiHa细胞30 min前加入NAC,进行预处理。使用CellTiter 96 AQueous单溶液细胞增殖检测试剂盒检测细胞存活率;利用2′,7′－二氯荧光黄双乙酸盐（ DCFH－DA）进行荧光探针标记测定胞内ROS水平;免疫印迹法检测cleaved caspase－3和PARP－1蛋白表达。结果显示, NAC预处理可降低Tv ESP的细胞毒性作用。 Tv ESP作用SiHa细胞16 h后,细胞内ROS水平升高,但经过NAC预处理后胞内ROS生成显著降低。此外, NAC预处理后能显著下调Tv ESP诱导的cleaved caspase－3和cleaved PARP－1的表达水平。     

阴道栓剂治疗阴道毛滴虫感染小鼠的实验研究

本实验以小白鼠为实验动物，作阴道毛病虫感染的动物模型，对阴道栓剂抗鼠体内阴道毛滴虫疗效进行了研究，实验分：配伍栓剂组、甲硝唑原粉组、环丙沙星原粉组和１％ＳＣＣ对照组。感染后２４ｈ采用一次性灌胃给药，１０ｄ后解剖小鼠，观察脓肿形成及愈合，镜检腹水和脓液。结果显示：栓剂治疗组滴虫清除率为５０％和９０％时，其剂量低于甲硝唑原份治疗剂量，两者比较具有显著性差异（Ｐ＜０．０５），而栓剂中杀淋球菌药环丙沙星原粉单独使用不能杀灭阴道毛滴虫。可见，甲硝唑与环丙沙星联合应用产生了一定的协同作用，减少了甲硝唑原粉杀滴虫的用量。     

香薷体外抗阴道毛滴虫的透射电镜观察

目的探讨香薷体外杀灭阴道毛滴虫的作用机制。方法将1∶4浓度的香薷水提液作用于体外培养的阴道毛滴虫,并采用透射电镜观察香薷作用2和4h后毛滴虫的超微结构变化。结果香薷作用2h后毛滴虫粗面内质网排列紊乱并脱颗粒,多聚核糖体解聚,高尔基复合体肿胀变形,胞质内空泡增多,核膜不完整,核质变疏松;作用4h后,胞膜破损,内容物外溢,内部结构消失,虫体死亡。结论香薷具有较强的抗滴虫作用,可破坏阴道毛滴虫的内膜系统。     

千里光提取物体外抗阴道毛滴虫的效果观察

目的研究千里光提取物对体外培养的阴道毛滴虫的杀虫效果。结论体外培养阴道毛滴虫,选择千里光提取物进行体外抗阴道毛滴虫实验,千里光药液浓度经稀释后分别为4,2,1,0.5,0.25mg/ml。同时以甲硝唑为阳性对照,乙醇及RPMI-1640为阴性对照。结果体外实验表明千里光在24h时杀灭阴道毛滴虫的最低有效浓度为2mg/ml,其杀虫效果与甲硝唑近似;阴道毛滴虫在RPMI-1640培养液中生长良好,增值3倍。结论千里光提取物在体外具有杀灭和抑制阴道毛滴虫生长的作用。     

抗阴道毛滴虫单克隆抗体在临床诊断中的应用

用抗阴道毛滴虫单克隆抗体,结合到聚苯乙烯胶乳微粒上。该胶乳试剂可以与滴虫的可溶性和不溶性抗原发生特异性凝集,对147份阴道拭子标本,分别用湿片直接镜检和胶乳凝集试验(LAT)进行比较,LAT阳性检出率比湿片镜检法略高(28％比18％)。LAT方法简便,3～5分钟可得出结果,与其它阴道炎分泌物没有凝集反应。表明LAT是一种敏感、特异、简单、快速的方法。     

阴道毛滴虫封存标本制作方法的改进

目的探索改进阴道毛滴虫封存标本的制作方法。方法取附属医院妇科门诊阴道毛滴虫患者的阴道分泌物,在无菌条件下接种于改良的肝浸汤培养基中,加入2ml无菌健康马血清及5×10^4U/ml青霉素,置37℃培养箱内培养,选取转种3代培养36h的阴道毛滴虫,采用载玻片的包装隔离纸带拖片,经改进的染液染色制片。结果封片标本中虫体分布均匀,伸展好。细胞浆呈淡蓝色,波动膜、轴柱、鞭毛呈蓝色,核红色,核仁紫红色。结论采用本方法制作的阴道毛滴虫封片标本虫体结构清晰,形态特征明显。     

成熟外周T淋巴细胞激活诱导细胞死亡

通过ＣＤ３－ＴＣＲ途径介导的信号，能在不成熟胸腺细胞和Ｔ细胞杂交瘤诱导出程序细胞死主恨。Ｄｉｅｔｅｒ Ｋａｂｅ１ｉｓｚ和同事收集了最近积累的关于在外周成熟Ｔ细胞也能诱导出程序细胞死亡的证据，这可能为细胞免疫调节的一个重要方面。     

两种快速染液对阴道毛滴虫染色效果的比较

本文比较了两种快速染液对阴道毛滴虫(Trichomonas Vaglnalis)的染色效果。结果表明，新疆研制的快速染液染色速度快、着色均匀、阴道毛滴虫的结构清晰、核浆对比突出。操作和观察省时省力，对于大面积的妇科病普查具有很高的优越性。     

山西长治市妇科门诊病人阴道毛滴虫感染流行病学调查

目的对长治市妇科门诊病人阴道毛滴虫感染情况进行调查，分析其流行因素，为制定本地区阴道毛滴虫防治措施提供科学依据。方法随机选择2007年12月至2008年11月在长治市晋城妇幼保健医院妇科门诊就诊的妇女作为调查对象，用阴道分泌物生理盐水涂片法检查阴道毛滴虫，并对受检者年龄、职业、文化水平、性行为等进行问卷调查。结果共检测516份标本，55份阳性，滴虫阳性率为10．66％。对所收集资料进行统计学分析得出，阴道毛滴虫感染与年龄、职业、文化水平、性病知识知晓水平、性行为等因素具有相关性。结论阴道毛滴虫感染是该地区妇科门诊患者的主要病原体之一；应根据其流行因素，制定预防保健措施，从保障该地区妇女生殖健康。     

The presence of dsRNA virus in Trichomonas vaginalis isolates from symptomatic and asymptomatic Indian women and its correlation with in vitro metronidazole sensitivity

Purpose: Trichomonas vaginalis, a protozoan parasite, is the causative agent of human trichomoniasis, the most common non-viral sexually transmitted disease. The infection encompasses from a complete asymptomatic presentation to severe sequelae; yet, the virulence markers have been poorly understood. It is suggested that the presence of Trichomonas vaginalis virus (TVV) in T. vaginalis may have an impact on its virulence, and its relatedness to in vitro metronidazole resistance has been reported. The aim of the study was to assess the presence of TVV in fresh and Long –Term Cultivated (LTC) maintained T. vaginalis isolates from symptomatic (S) and asymptomatic (AS) Indian women and its relatedness, if any, with symptomatology and in vitro drug sensitivity. Materials and Methods: One thousand women (537 S and 463 AS) were screened for the presence of T. vaginalis by wet smear and culture examination of vaginal swab and urine sample. Fresh and LTC (6 months–2 years) maintained 15 isolates each from 15 S and 15 AS women were subjected to agarose gel electrophoresis following total cellular RNA extraction to evaluate the presence of double stranded (ds) RNA viral infection. The susceptibility of isolates to metronidazole was determined in vitro. Results: On agarose gel electrophoresis, three bands (5.5, 2.5 and 1.5 kb) were observed in all the 30 fresh isolates from 15 S and 15 AS women and only in 7 LTC isolates from 3 S and 4 AS women. All the fresh isolates harbouring TVV were found to be sensitive to metronidazole in vitro irrespective of the symptomatology of subjects, and out of seven LTC isolates harbouring TVV, six were sensitive to metronidazole and one showed borderline resistance. Conclusions: The results suggest that the presence of TVV alone may not be a virulence marker and loss of TVV on LTC appears to be related to drug resistance. The T. vaginalis Indian isolates are sensitive to metronidazole.     

奥硝唑不同剂型抗阴道毛滴虫作用的比较研究

目的研究奥硝唑原料药、凝胶剂和栓剂在体内外抗阴道毛滴虫的作用。方法体内实验：在小鼠阴道内连续多日注入不同浓度奥硝唑、奥硝唑凝胶或奥硝唑栓剂和虫体,停药1 d后取阴道灌洗液涂片观察有无活动的阴道毛滴虫。体外实验：将阴道毛滴虫培养后分别接种到在含不同浓度奥硝唑、奥硝唑凝胶和奥硝唑栓剂的细胞培养板孔中,于不同时间在倒置显微镜下观察虫体形态变化,计数死虫百分率。结果体内试验中连续5 d注入药物和虫体的奥硝唑原料药组、奥硝唑凝胶剂组、奥硝唑栓剂组镜检阳性率均为0（0/20）,而无药对照组阳性率为100%（20/20）,差异有统计学意义（P〈0.01）。体外实验中药物作用滴虫24 h后,各种药物的最低有效抑虫浓度（虫体100%死亡）依次为：奥硝唑凝胶0.0025 mg/ml,奥硝唑原料0.0049 mg/ml,奥硝唑栓剂0.0195 mg/ml。结论奥硝唑凝胶剂具有良好的体内外抗阴道滴虫作用,其作用优于奥硝唑栓剂。     

Detection of Trichomonas vaginalis in endocervical and ectocervical smears

The objective of this study was to determine the relative sensitivity of detecting Trichomonas vaginalis in endocervical as compared to ectocervical Papanicolaou-stained smears. The average number of organisms in 10 (200×) high-power fields (hpf) per smear was used for comparing the presence of trichomonads in the two smear types. In every pair of ectocervical-endocervical smears, there were always more trichomonads in the ectocervical smear. Fifty percent of cases showed at least 25 more trichomonads per 200× hpf in the ectocervical smear than in the endocervical smear. In 20 of 50 cases (40%), trichomonads were present only in the ectocervical smear. The organisms were found only in the endocervical smears of women with a heavier ectocervical trichomonad burden. In conclusion, trichomonads are detected in the Papanicolaou-stained ectocervical smear more often than in the endocervical smear; therefore, examination of an adequate ectocervical smear has greater utility than the endocervical smear for the diagnosis of trichomoniasis. Diagn Cytopathol 1996;14:273–276. © 1996 Wiley-Liss, Inc.     

Identification of weak points prone for mutation in ferredoxin of Trichomonas vaginalis

Trichomonas vaginalis, the causative agent for human trichomoniasis, is a problematic sexually transmitted disease mainly in women. At present, metronidazole-resistant trichomoniasis is an infrequent but challenging problem with no universally successful treatment. Genetic mutation is believed to be an important factor leading to increasing drug resistance. Understanding the mutation status will help to design accurate strategies of therapy against mutant strains of T. vaginalis. The author performed a bioinformatic analysis to determine positions that tend to comply peptide motifs in the amino acid sequence of ferredoxin of T. vaginalis. Based on this study, the weak linkages in the studied protein can be identified and can be useful information for prediction of possible new mutations that can lead to drug resistance. In addition, the results from this study can be good information for further research on the diagnosis for mutants and new effective drug development.     

一个含有25bp内含子的阴道毛滴虫Rabl-like新基因

目的 克隆和鉴定一个新的阴道毛滴虫Rabl-like基因（TvRabl-like）及其内含子。方法 我们从一阴道毛滴虫cDNA表达文库中分离出一个cDNA克隆，它与各物种的Rab家族蛋白有较高的同源性，因此我们进一步用BLASTP、RPS-BLAST、ClustalW和MEGA3等分析软件对该cDNA克隆进行了序列分析和进化树分析；用PCR和RT-PCR等技术分别对该基因组和mRNA进行了扩增和测序分析。结果 序列分析结果表明该eDNA克隆长705bp，开放阅读框具603bp，推测肽链含有200个氨基酸。序列比较分析结果提示该cDNA克隆所推测的蛋白质是一个Rabl亚家族的亚型。进化树分析也表明它属于阴道毛滴虫Rabl亚家族。基因组PCR扩增和测序分析表明该基因包含一个25bp的内含子，该内含子具有阴道毛滴虫和其它真核生物较大内含子所具备的典型的5’GT-AG-3’和分支位点基序。RT-PCR产物及其测序分析表明在该基因的转录本中存在着未剪切和剪切后的mRNA，说明确实有内含子的存在。结论 TvRabl-like基因属于阴道毛滴虫Rabl亚家族，该基因含有一个25bp的内含子。该内含子是至今发现的最小的阴道毛滴虫基因内含子之一，很可能也是真核生物中最小的内含子。对诸类最低等真核生物内含子的研究将有助于我们理解真核生物内含子的起源和进化。     

Trichomonas vaginalis P16 Immunoreactivity in cervicovaginal Pap tests: a diagnostic pitfall

Detection of p16 is emerging as a useful biomarker for Human Papillomavirus (HPV)-related dysplastic and malignant lesions of the cervix, and as such has potential application to cervicovaginal Papanicolaou (Pap) specimens. While evaluating p16 immunocytochemistry in our laboratory we observed one Pap test in which Trichomonas vaginalis stained positively for p16. We therefore proceeded to determine the frequency of T. vaginalis immunoreactivity for p16 in 10 consecutive, satisfactory, liquid-based Pap tests diagnosed as negative for intraepithelial lesion or malignancy in which T. vaginalis was present. For each case, a ThinPrep slide prepared from residual vial material was immunostained with p16. In an additional case, a prepared cell block was stained with p16. T. vaginalis were consistently p16 positive. We document, for the first time, p16 immunoreactivity of T. vaginalis that, on a Pap slide and cell block, may be morphologically misinterpreted as small dysplastic or malignant epithelial cells. The presence of this parasite in Pap tests may also potentially hinder the use of p16 as an adjunct to liquid-based cervical screening cytology.     

Clinical and laboratory evidence of Trichomonas vaginalis infection among women of reproductive age in rural area

Background: Vaginitis is a commonly encountered complaint and one of the most frequent reasons for patient visit to obstetrician-gynaecologists. Three vaginal infections are frequent causes of a vaginal discharge: (1) bacterial vaginosis, (2) vulvovaginal candidiasis and (3) trichomonas vaginitis. Differences in the clinical presentation are helpful in diagnosis. Characteristic signs and symptoms for these three vaginal infections are distinct, but on many occasions, they are overlapping. The aim of the present study was to find the prevalence and correlation between the clinical spectrum and laboratory evidence of Trichomonas vaginalis infection by simple, reliable, confirmatory and specific method, i.e. microscopic examination of wet mount preparation and acridine stain of vaginal fluid. Materials and Methods: Irrespective of HIV status, a total of 156 women with vaginal discharge were studied for establishing diagnosis of genital tract infection. The cases of bacterial vaginosis and vulvovaginal candidiasis were excluded from the study. Vaginal speculum assisted high vaginal swabs were collected from women with discharge, during collection vagina was inspected for obvious signs. Results: Of the 156 women with vaginal discharge, 19 (12.06 %) showed T. vaginalis infection. All the women belonged to active reproductive age group, i.e. 20-40 years. Itching dysuria, and offensive, malodorous, thin, yellowish vaginal discharge were the main and consistent complaints. Only in 2 (1.52%) cases, vaginal speculum examination revealed erythema and punctuate haemorrhage, the so-called "strawberry' vagina. The pH was recorded to be >4.5. Conclusion: Clinical differentiation of various forms of infectious vaginitis is unreliable. The prevalence of T. vaginalis infection at 12.06% was found among rural young women of reproductive age using simple and reliable screening wet mount microscopy.